青海海北地区牦牛病毒性腹泻病毒流行病学调查及5'-UTR遗传进化分析

doi:10.16431/j.cnki.1671-7236.2025.03.026

Abstract

Abstract: 【Objective】 The aim of this study was to understand the prevalence and molecular characteristics of Bovine viral diarrhea virus (BVDV) in yaks in Haibei,Qinghai province.【Method】 Viral RNA was extracted from 148 diarrhea yak feces samples collected from Haibei,Qinghai province,and cDNA was synthesized by reverse transcription.BVDV 5'-UTR conserved region was amplified with the template,and a Real-time quantitative PCR method was established to detect BVDV infection in yak farms in Haibei,Qinghai province.4 BVDV positive samples were selected for PCR amplification and sequencing of the BVDV 5'-UTR region sequence,similarity comparison with different subtypes of BVDV and strains of the same genus in NCBI,and genetic evolution analysis was conducted based on the BVDV 5'-UTR region.【Result】 In this study,Real-time quantitative PCR detection method was successfully established based on BVDV 5'-UTR.The correlation coefficient (R²) of standard curve was 0.997,and the linear relationship was good when the standard concentration was 2.69×10²-2.69×10⁹ copies/μL,and the minimum detection limit was 2.69×10² copies/μL.The results of repeatability test showed that the coefficient of variation was less than 1.7%,which had good repeatability and stability.Yak diarrhea feces samples from Menyuan,Qilian,Haiyan and Huangyuan counties in Haibei area of Qinghai province were detected by Real-time quantitative PCR method，the positive rates of BVDV were 45.00%,36.00%,30.77% and 29.00%,respectively.The results of BVDV 5'-UTR sequence comparison showed that the 4 yak BVDV isolates were clustered in the same independent branch with the BVDV-1a subtype strains from Iran and United States,and the nucleotide sequence similarity was 76.6%-99.7%.【Conclusion】 This study identified BVDV-1a as one of the important pathogens causing yak diarrhea in Haibei area of Qinghai province,and provided scientific basis for yak farms in this area to take timely and effective prevention and control measures.At the same time,the molecular epidemiological data of local BVDV were enriched,which provided a theoretical basis for the prevention and control of bovine viral diarrhea disease and vaccine research and development in this area.

Key words: yak; Bovine viral diarrhea virus (BVDV); epidemiology; Real-time quantitative PCR; genetic evolution analysis

CLC Number:

S852.65⁺3

LIN Weishan, MA Doudou, LEI Mengtong, WEI Bin, LI Guocai, WANG Guanghua, WANG Geping, JIAN Yingna, LI Xiuping. Epidemiological Investigation and Genetic Evolution Analysis of 5'-UTR of Bovine Viral Diarrhea Virus in Yaks in Haibei，Qinghai Province[J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(3): 1241-1249.

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Epidemiological Investigation and Genetic Evolution Analysis of 5'-UTR of Bovine Viral Diarrhea Virus in Yaks in Haibei，Qinghai Province

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